〔Abstract〕 Objective To investigate the impact of SOX4 on ovarian granulosa cells, stable overexpression of SOX4 was achieved in human KGN cell line, followed by analysis of its effects on proliferation, migration and apoptosis. Methods The recombinant lentiviral plasmid pLV-EF1a-GFP/Puro-SOX4 was generated through homologous recombination with linearized pLV-EF1a-GFP/Puro vector.Human ovarian granulosa cells(KGN cell line) were transduced with Lentiviral expression vectors.KGN cells infected with pLV-EF1a-GFP/Puro-NC were served as the LV-CON group, while those infected with pLV-EF1a-GFP/Puro-SOX4 were designated as the LV-SOX4 group.Following transfection, puromycin selection was employed to establish stable SOX4-expressing KGN cells.The expression levels of SOX4 m RNA and protein in KGN cells from the LV-CON and LV-SOX4 groups were assessed using RT-qPCR and Western blot analysis.Cell proliferation was assessed using the CCK-8 assay in both LV-CON and LV-SOX4 groups.Cell migration ability was evaluated by means of a cell scratch test in these two groups.The proportion of apoptotic cells was determined via flow cytometry analysis in both LV-CON and LV-SOX4 groups. Results The sequencing results of pLV-EF1a-GFP/Puro-SOX4 indicated a complete match between the inserted gene sequence and the SOX4 mRNA sequence.The lentiviral titers were 7×108TU/ml in the LV-CON group and 1×108TU/ml in the LV-SOX4 group.The recombinant plasmid was successfully transfected into KGN cells with a transfection efficiency of over 90% under fluorescence inverted microscopy.The results of RT-qPCR and Western blot tests demonstrated a significant increase in the expression level of SOX4 in KGN cells of LV-SOX4 group compared to that of LV-CON group(t=3.10,P<0.05;t=14.20,P<0.05).The CCK-8 assay results demonstrated that the LV-SOX4 group exhibited a significant increase in cell proliferation(24 h:t=45.92,P<0.01; 72 h:t=25.60,P<0.01) compared to the LV-CON group.The cell scratch assay indicated that the migratory capacity of KGN cells in the LV-SOX4 group was significantly enhanced(t=7.65,P<0.01) compared to that in the LV-CON group.The LV-SOX4 group exhibited a significant reduction in apoptosis ratio(t=25.84,P<0.01) compared to the LV-CON group. Conclusion SOX4-overexpressing KGN cell line was successfully established, and the overexpression of SOX4 facilitated proliferation and migration while inhibiting apoptosis in human ovarian granulosa cells.